High performance mass spectrometry at the University of Birmingham

Mass spectrometry is well-established as a technique for characterisation of molecular structure. The chief reason for the success of mass spectrometry is that every molecule has a mass, unlike other properties sometimes measured in bioanalytical chemistry, such as a magnetic moment or absorbance of light at a particular wavelength. The mass of a molecule depends on the composition of its atoms, as each atom has a unique mass. As long as the accuracy of the mass spectrometer is high enough, then measurement of a molecule's mass allows determination of the elemental composition. The more complex the sample, either in terms of the number of components or in terms of the size of the molecule (the number of atoms), the more challenging is mass analysis. This fact is due to overlapping signals from different molecules or versions of the same molecule (isotopomers) and in order to address the problem, a very high resolution mass spectrometer is required.

Biological research is overflowing with sample types that require high resolution mass spectrometry: Metabolomic samples contain thousands of low molecular weight species. Proteins are large molecules associated with a vast array of chemical modifications, and which form loosely-bound complexes with themselves, other proteins and other molecule types. The aim of this proposal is to make available the latest developments in the highest performance type of mass spectrometry - Fourier transform ion cyclotron resonance mass spectrometry - to biological research at the University of Birmingham and beyond. The proposal brings together a broad range of researchers from diverse disciplines to address several key strategic priorities for BBSRC (Healthy ageing across the life course, Synthetic biology, Combatting antimicrobial resistance, Food, nutrition & health, Technology for the biosciences).

The aim of the proposal is to purchase a Bruker Solarix 7 Tesla Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. The instrument is equipped with the latest in ICR cell design (the Paracell) which offers huge improvements in resolving power over previous designs. The University of Birmingham has been at the centre of FT-ICR applications research for the past decade. Our current instrument, which was state-of-the-art at time of purchase, offers a maximum resolving power of 1,000,000 at m/z 400 and mass accuracy of 1 ppm. The proposed instrumentation offers a maximum resolving power is 10, 000, 000 at m/z 400 - a tenfold improvement - and a mass accuracy of 600 ppb. The instrument also incorporates the latest in ion funnel design to allow transport of fragile assemblies such as protein complexes. To date, over 100 research groups have benefitted from access to FT-ICR mass spectrometry though the Advanced Mass Spectrometry Facility at the University of Birmingham (internal, national, international and industry). The vastly enhanced features of the latest generation of FT-ICR mass spectrometer will enable us to make a step-change in terms of information available (numbers and nature of metabolites, high molecular weight proteins and protein complexes, protein post-translational modifications) to researchers in the biological sciences.

Who will benefit from the research?
The proposal aims to exploit the power of high performance mass spectrometry across of broad range of disciplines within the biological sciences. The beneficiaries of this work will be the pharmaceutical industry, the agri-food industry, the NHS and its patients, biotechnology industry, analytical instrument manufacturers.

How will they benefit from the research?
The proposed research has the potential to contribute significantly to the nation's health and wealth:

Agri-food industry: Campylobacter jejuni remains a major foodborne pathogen, largely originating from intestinal carriage of the organism in farm animals, especially poultry. Control of this organism to reduce its incidence and resulting economic losses is a major challenge to industry. The proposed research will address this challenge by providing insight into the attributes of the organism that determine its fundamental fitness and ability to withstand measures designed to eliminate it from food production systems.
Similarly, understanding how host attachment leads to activation of enterohemorrhagic E. coli (EHEC) virulence genes will enable us to devise small molecules to interfere with this pathway and thus stop colonization of food animals and disease in humans. Novel ways to stop colonization with EHEC and other food-borne pathogens will be of immense value to the agri-food industry and help ensure our supply with safe and healthy food in the future.

Pharmaceutical industry: Novel ways to treat EHEC infections in humans are sought after, as antibiotics are counter-indicated in EHEC infections as they often trigger the production of shiga toxins, which lead to severe complications such as hemolytic uremic syndrome and increased morbidity and mortality. The research will ultimately help to replenish the drug development pipeline with novel small molecules, which will alleviate some of the problems we have encountered with antibiotics.
The lipidomics and proteomics study of protein YraP will also benefit the pharmaceutical industry. Henderson has already established collaborations with Novartis on the topic of this project, speicifically seeking to understand the molecular basis of the adjuvanticity of YraP in the Bexsero vaccine.

Biotechnology industry: Development of the bacteriophage/linear dichroism devices will benefit the biotechnology industry. Dafforn has established a spin-out company based on the technology, taking advantage of a BBSRC Enterprise Fellowship.

Analytical instrumentation manufacturers: The work will enable the development of new workflows which can be employed in a broad range of new biological applications. Through the proposed research, these new applications will lead to increased sales this fostering global economic performance and the economic competitiveness of the UK.